Team:Kyoto/GSDD/Notebook/0817-0823

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0727-0802
Construction.
0803-0809
Construction.
0810-0816
Construction.
0817-0823
Construction.PCR.
0824-0830
Construction.PCR.
0831-0906
Construction.PCR.MPR.
0907-0913
Construction.PCR.MPR.
0914-0920
Construction.PCR.MPR.
0921-0927
Construction.PCR.MPR.
0928-1004
Construction.PCR.MPR.
1005-1011
Construction.PCR.MPR.
1012-1018
Construction.MPR.Observation.

0817-0823 : Construction.PCR.


Tuesday, 18 August

To Do

  • PCR
  • Restriction Enzyme Digestion
  • Electrophoresis
  • Gel Extraction kit
    • 3-O-2
    • pSB1A2

Results

3-O-2
primerF
AACTGCAGAACCAATGCATTGGagaacgtctctcactgaac
primerR
aaaattttGGACTAGTCCttgaatcttagcccccttg

Kyoto 0818 1.jpg

  • Length
3-O-2+BBa_J63010
6103bp
3-O-2 PCR product
2851bp

We can not find the bands of PCR products. So, we will do PCR again, changing the volume of MgSO4, 3ul, 4.5ul, 6ul.

pSB1A2

Kyoto 0818 2.jpg

  • Length
pSB1A2
2079bp
conc.
sample nameconc./(ng/ul)
pSB1A2(EcoR1,Spe1)9.5
pSB1A2(Xba1,Pst1)5.6

We use pSB1A2 in order to insert PCR products and sequence them. But the concentrations of this sample are very low. So, we will try again.



Friday, 21 August

To Do

  • PCR
  • Restriction Enzyme Digestion
  • Electrophoresis
  • Gel Extraction kit
    • 3-O-2
    • pSB1A2


Results

3-O-2
primerF
AACTGCAGAACCAATGCATTGGagaacgtctctcactgaac
primerR
aaaattttGGACTAGTCCttgaatcttagcccccttg
  • Length
3-O-2+BBa_J63010
6103bp
3-O-2 PCR product
2851bp
pSB1A2
  • Length
pSB1A2
2079bp
conc.
sample nameconc./(ng/ul)
pSB1A2(EcoR1,Spe1)13.4
pSB1A2(Xba1,Pst1)11.9

We use pSB1A2 in order to insert PCR products and sequence them.